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Fig. 1
Cftr PDZBD but not its channel function is required for its interaction with Dvl2 and hematopoiesis
a Schematic drawing of PDZBD in Cftr and PDZ domain in Dvl2. Referring to Gee, H. Y. (2011)[57] and Wallingford, J. B. (2005)[58]. NBD nucleotide-binding domain, TM transmembrane domain, R regulatory domain, DIX Dishevelled and AXIN domain, DEP Dishevelled EGL-10 Pleckstrin domain. b Co-immunoprecipitation (Co-IP) of endogenous Dvl2 and Cftr in zebrafish embryos. c Co-localization of Cftr and Dvl2 in zebrafish embryo and HEK293 cells. Arrowheads indicate co-localized sites. Scale bar 5 μm. d In vitro binding assay identifies the physical interaction of Cftr with Dvl2. e Co-IP of exogenous Cftr and Dvl2 in HEK293 cells showing lack of interaction when Cftr PDZBD or Dvl2 PDZ domain is deleted. f Co-IP showing the interaction of Dvl2 with G551D, but not Cftr PDZBD mutants (either deletion or point mutation), and western blotting showing that Cftr and G551D overexpression increased the expression of Dvl2 and active-β-catenin more significantly than that induced by Cftr PDZBD mutants. g Immunofluorescence analysis in HEK293 cells shows that either of the CFTR PDZBD mutants is not co-localized with Dvl2, whereas overexpressed G551D is co-localized with Dvl2. h Effects of different cftr mutants mRNA in rescuing hematopoietic defects in cftr mutant zebrafish embryos. Injection of cftr PDZBD mutants individually, with deletion or point mutation, could not rescue the hematopoietic defect in cftr mutant embryos. Injection of G551D rescues the hematopoietic defect in cftr mutant embryos. Embryos shown are dorsal views with anterior to the top at 8-somite stage (13 hpf)